pFastBac CT-TOPO (linearized)

Linearized baculovirus expression vector for TOPO® cloning of blunt PCR products and addition of a cleavable C-terminal 6xHis tag.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsInsect Cell VectorspFastBac CT-TOPO (linearized)
BstZ17I (4590) AccI (4589) SnaBI (4587) BbsI (4463) BsrGI (4116) PflFI - Tth111I (3934) BsmBI - Esp3I (3888) BseRI (3667) EcoRV (3531) SacII (3474) BstXI (3423) MscI (3415) BspQI - SapI (3085) PspFI (2668) BseYI (2664) AlwNI (2559) BamHI (4738) AgeI (4744) End (4754) Start (0) 6xHis PaeR7I - PspXI - XhoI (49) SphI (59) Acc65I (61) KpnI (65) HindIII (67) MfeI (182) HpaI (195) BclI * (331) AvrII (346) NgoMIV (833) NaeI (835) BanII (867) BsaHI (1540) PvuI (1711) FspI (1857) BpmI (2011) pFastBac™/CT-TOPO® 4754 bp
BstZ17I  (4590)
1 site		 G T A T A C C A T A T G
AccI  (4589)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SnaBI  (4587)
1 site		 T A C G T A A T G C A T
BbsI  (4463)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BsrGI  (4116)
1 site		 T G T A C A A C A T G T
BsrGI is typically used at 37°C, but is even more active at 60°C.
PflFI  (3934)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3934)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsmBI  (3888)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (3888)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
BseRI  (3667)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
EcoRV  (3531)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
SacII  (3474)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
BstXI  (3423)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
MscI  (3415)
1 site		 T G G C C A A C C G G T
BspQI  (3085)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (3085)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PspFI  (2668)
1 site		 C C C A G C G G G T C G
BseYI  (2664)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (2559)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
BamHI  (4738)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
AgeI  (4744)
1 site		 A C C G G T T G G C C A
End  (4754)
0 sites
Start  (0)
0 sites
PaeR7I  (49)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (49)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (49)
1 site		 C T C G A G G A G C T C
SphI  (59)
1 site		 G C A T G C C G T A C G
Acc65I  (61)
1 site		 G G T A C C C C A T G G
KpnI  (65)
1 site		 G G T A C C C C A T G G
HindIII  (67)
1 site		 A A G C T T T T C G A A
MfeI  (182)
1 site		 C A A T T G G T T A A C
HpaI  (195)
1 site		 G T T A A C C A A T T G
BclI  (331)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
AvrII  (346)
1 site		 C C T A G G G G A T C C
NgoMIV  (833)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (835)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
BanII  (867)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
BsaHI  (1540)
1 site		 G R C G Y C C Y G C R G
BsaHI is typically used at 37°C, but is even more active at 60°C.
PvuI  (1711)
1 site		 C G A T C G G C T A G C
FspI  (1857)
1 site		 T G C G C A A C G C G T
BpmI  (2011)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AmpR
1293 .. 2153  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1293 .. 1361  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1293 .. 2153  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1362 .. 2153  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1293 .. 2153  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2324 .. 2912  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2324 .. 2912  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
GmR
3508 .. 4041  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
3508 .. 4041  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
708 .. 1162  =  455 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
708 .. 1162  =  455 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
Tn7R
3217 .. 3441  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
3217 .. 3441  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
359 .. 524  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
359 .. 524  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
196 .. 330  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
196 .. 330  =  135 bp
SV40 polyadenylation signal
AmpR promoter
1188 .. 1292  =  105 bp
AmpR promoter
1188 .. 1292  =  105 bp
polyhedrin promoter
4610 .. 4701  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4610 .. 4701  =  92 bp
promoter for the baculovirus polyhedrin gene
TEV site
7 .. 27  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
7 .. 27  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
6xHis
28 .. 45  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
28 .. 45  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
Pc promoter
4230 .. 4258  =  29 bp
3 segments
   Segment 3:  -10  
   4230 .. 4235  =  6 bp
class 1 integron promoter
Pc promoter
4230 .. 4258  =  29 bp
3 segments
   Segment 2:  
   4236 .. 4252  =  17 bp
class 1 integron promoter
Pc promoter
4230 .. 4258  =  29 bp
3 segments
   Segment 1:  -35  
   4253 .. 4258  =  6 bp
class 1 integron promoter
Pc promoter
4230 .. 4258  =  29 bp
3 segments
class 1 integron promoter
ORF:  3646 .. 3978  =  333 bp
ORF:  110 amino acids  =  12.0 kDa
ORF:  4252 .. 4494  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  1293 .. 2153  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3351 .. 3653  =  303 bp
ORF:  100 amino acids  =  11.8 kDa
ORF:  4143 .. 4622  =  480 bp
ORF:  159 amino acids  =  18.5 kDa
ORF:  4233 .. 4541  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  1757 .. 2023  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3508 .. 4041  =  534 bp
ORF:  177 amino acids  =  19.4 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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